P-706 Non-invasive analysis of mitochondrial DNA mutation for embryo evaluation.

Abstract Study question Is it possible to amplify whole mitochondrial DNA (mtDNA) and detect all mtDNA mutation in human embryo from spent media after culture? Summary answer We were able 43% of medium 24-hour culture. The most could be detected. What is known already Preimplantation genetic testing for aneuploidy (PGT-A) with biopsy now commonly used improve pregnancy rate. However, invasive requires highly trained embryologist induced higher risk pre-eclampsia. Therefore, non-invasive methods cell-free have been studied. Our group previously showed euploid or viable embryos at post-implantation stage lower number non-synonymous mutations this may help select the best blastocyst transfer. Although reported contain mtDNA, remains unclear whether can assessed DNA. design, size, duration Donated low grade blastocysts cultured additional 24 hours new media, obtained culture (non-invasive samples). For comparison, trophectoderm cells biopsied each (invasive sample). selective amplifications done both samples. performed sequencing by next generation (NGS) compared between samples clarify capability assessment mutation. Participants/materials, setting, Under ethical review Yokohama City University informed consent patients, we collected which discarded because slow development morphologically low-grade. extracted selectively amplified long-range PCR two overlapping amplicons. NGS was successfully mtDNA. 90% more heteroplasmy level investigated. Main results role chance 34 13 patients. All individually 20 μl drop newly prepared media. Amplified visualized presence 8 kb band following agarose gel-apheresis fragments PCR. Both detected 41% (14/34) Only one fragment 14% (5/34) completely corresponding NGS. In results, identified these also location matched 78.5% (11/14) embryos, other 3 numbers even though no existed This means, if amplification completed, evaluated a probability about 80% but include 20% false positive mutations. To realize this, need method. Limitations, reasons caution only clinically restriction. And necessary 24-hours obtain are co-cultured until they donated. Because small available, keep collecting accurate evaluation. Wider implications findings If analyzed medium, high-quality selected without complicated procedure. analysis clinical transfer, contribute further improvement rate any invasion embryo. Trial registration Japan Society Promotion Science (JSPS) KAKENHI Grant-in-Aid Scientific Research C (Grant Number JP21K09474) Early-Career Scientists JP20K18169)